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| United States Patent |
5,300,279
|
|
Simon
, et al.
|
*
April 5, 1994
|
Organic amine phosphonic acid complexes for the treatment of calcific
tumors
Abstract
Particle-emitting radionuclides, e.g. Gd-159, Ho-166, Lu-177 and Yb-175,
have been complexed with organic aminoalkylenephosphonic acids. These
complexes have been found useful in compositions for the therapeutic
treatment of calcific tumors or the relief of bone pain in animals.
| Inventors:
|
Simon; Jaime (Angleton, TX);
Garlich; Joseph R. (Lake Jackson, TX);
Goeckeler; William F. (Midland, MI);
Wilson; Davis A. (Richwood, TX);
Volkert; Wynn A. (Columbia, MO);
Troutner; David E. (Phoenixville, PA)
|
| Assignee:
|
The Dow Chemical Company (Midland, MI)
|
| [*] Notice: |
The portion of the term of this patent subsequent to November 19, 2008
has been disclaimed. |
| Appl. No.:
|
629894 |
| Filed:
|
December 19, 1990 |
| Current U.S. Class: |
424/1.77; 534/10 |
| Intern'l Class: |
A61K 043/00 |
| Field of Search: |
424/1.1
534/10
|
References Cited
U.S. Patent Documents
| 3851044 | Nov., 1974 | Adler et al. | 424/1.
|
| 3852414 | Dec., 1974 | Adler et al. | 424/1.
|
| 3931396 | Jan., 1976 | Bardy et al. | 424/1.
|
| 3965254 | Jun., 1976 | Tofe et al. | 424/1.
|
| 3989730 | Nov., 1976 | Subramanian et al. | 424/1.
|
| 4017596 | Apr., 1977 | Loberg et al. | 424/1.
|
| 4058704 | Apr., 1978 | Simon et al. | 424/1.
|
| 4075314 | Feb., 1978 | Wolfangel et al. | 424/1.
|
| 4104366 | Aug., 1978 | Schmidt-Dunker et al. | 424/1.
|
| 4399817 | Aug., 1983 | Benedict | 424/1.
|
| 4454106 | Jun., 1984 | Gansow et al. | 424/1.
|
| 4466951 | Aug., 1984 | Pittman | 424/1.
|
| 4500508 | Feb., 1985 | Srivastava et al. | 424/1.
|
| 4515767 | May., 1985 | Simon et al. | 424/1.
|
| 4560548 | Dec., 1985 | Simon et al. | 424/1.
|
| 4606907 | Aug., 1986 | Simon et al.
| |
| 4707352 | Nov., 1987 | Stavrianpoulos | 424/1.
|
| 4897254 | Jan., 1990 | Simon et al. | 424/1.
|
| 4898724 | Feb., 1990 | Simon et al. | 424/1.
|
| 5066478 | Nov., 1991 | Simon et al. | 424/1.
|
| Foreign Patent Documents |
| 1078731 | Jun., 1980 | CA.
| |
| 210043 | Jul., 1986 | EP.
| |
| 2109407 | Jun., 1983 | GB.
| |
Other References
Journal Nucl. Med., vol. 1 (1960) pp. 1-13.
Journal Nucl. Med., vol. 10 (1969) pp. 49-51.
Int. J. Clin. Pharmacol., 9, 3 (1974), pp. 199-205.
Journ. Nucl. Med., vol. 16 (1975) pp. 1080-1084.
Journal of Urology, vol. 116 (1976), pp. 764-768.
Seminars in Nucl. Med., vol. IX, No. 2 (Apr. 1979), pp. 114-120.
19th Int. Annual Meeting Soc. Nucl. Med. Europe, Bern, Switzerland Sep.
8/11, 1981 Journal Nucl. Med., vol. 24 (1983), p. P-125.
Journ. Nucl. Med., vol. 25 (1984) pp. 1356-1361.
Journ. Nucl. Med., vol. 25 (1984) p. P-129.
Int. J. Applied Rad. and Isotopes, Mathieu, L. et al. vol. 30, pp. 725-727
(1979).
Int. J. of Applied Rad. & Isotopes, vol. 14, pp. 129-135 (1963).
Technical Rept. from the Med. Dept., Brookhaven Nat. Lab. (BNL-24614).
Sixth Int. Symposium on Radiopharm. Chem. Paper No. 140 1986.
Sixth Int. Symposium on Radiopharm. Chem. Paper No. 141 1986.
Derwent Abst. 84-140663/23.
Chemical Abstract 87:179938h.
Chemical Abstract 91:97498h.
Chemical Abstracts 93:163556v.
Chemical Abstract 95:199803d.
Chem. Abst. 96:14590m.
Chem. Abst. 106:67404k.
|
Primary Examiner: Stoll; Robert L.
Assistant Examiner: Covert; John M.
Attorney, Agent or Firm: Kimble; Karen L.
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATION
This application is a continuation-in-part of our co-pending applications
Ser. No. 472,506 filed Jan. 30, 1990, now U.S. Pat. No. 5,066,478 which is
a divisional of Ser. No. 50,263 filed May 14, 1987, now U.S. Pat. No.
4,898,724, which is a continuation-in-part of Ser. No. 803,376 filed Dec.
4, 1985, now abandoned, which is a continuation-in-part of Ser. No.
738,010 filed May 28, 1985, now abandoned, which is a continuation-in-part
of Ser. No. 616,985, filed Jun. 4, 1984, now abandoned.
Claims
We claim:
1. A therapeutically effective composition comprising (1) an
aminophosphonic acid of the formula
##STR7##
wherein substituents A, B, C, D, E and F are independently selected from
hydrogen, methyl, ethyl, isopropyl, benzyl
##STR8##
and physiologically acceptable salts of the acid radicals and wherein X'
and Y' are independently hydrogen, methyl or ethyl radicals; n' is 2 or 3;
and m and m' each is independently 0-10; with the proviso that at least
one of said nitrogen substituents is the phosphorus-containing group of
Formula (B); and wherein R is
##STR9##
wherein X and Y are independently selected from hydrogen, hydroxyl,
carboxyl, phosphonic, and hydrocarbon radicals having from 1-8 carbon
atoms and physiologically acceptable salts of the acid radicals and n is
1-3, with the proviso that when n>1, each X and Y may be the same as or
different from the X and Y of any other carbon atoms; with the further
proviso that when m or m'.gtoreq.1 the E and F substituents may be same as
or different from any other substituent of any other nitrogen atom and
each R can be the same as or different from any other R and (2) at least
one radionuclide selected from G-159, Ho-166, Lu-177 and Yb-175.
2. The composition of claim 1 wherein the radionuclide is Ho-166.
3. The composition of claim 1 wherein the aminophosphonic acid has the
formula
##STR10##
wherein A, B and C are defined as in claim 1.
4. The composition of claim 3 wherein the aminophosphonic acid is
nitrilotrimethylenephosphonic acid or a physiologically acceptable salt
thereof and the radionuclide is Yb-175.
5. The composition of claim 1 wherein the aminophosphonic acid has the
formula
##STR11##
wherein m is zero and R is --CH.sub.2 CH.sub.2 -- and A, B, C, D, E, F and
m' are defined as in claim 1.
6. The composition of claim 5 wherein X and Y are hydrogen; n is 1; and X'
and Y' are each independently hydrogen, methyl or ethyl radicals.
7. The composition of claim 5 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid or a physiologically
acceptable salt thereof.
8. The composition of claim 7 wherein the radionuclide is Ho-166.
9. The composition of claim 5 wherein at least one of the nitrogen
substituents is
##STR12##
wherein X', Y' and n' are defined as in claim 1.
10. The composition of claim 9 wherein the aminophosphonic acid is
hydroxyethylethylenediaminetrimethylenephosphonic acid or a
physiologically acceptable salt thereof.
11. The composition of claim 10 wherein the radionuclide is Ho-166.
12. The composition of claim 5 wherein the radionuclide is Ho-166 and the
aminophosphonic acid is N-methylethylenediaminetrimethylenephosphonic
acid, N-isopropylethylenediaminetrimethylenephosphonic acid,
N-benzylethylenediaminetrimethylenephosphonic acid, or a physiologically
acceptable salt thereof.
13. The composition of claim 1 wherein the aminophosphonic acid is
##STR13##
wherein m is 1, m' is zero and R is --CH.sub.2 CH.sub.2 -- and A, B, C, D,
E, and F are defined as in claim 1.
14. The composition of claim 13 wherein X and Y are each hydrogen; and n is
1; and X' and Y' are each independently hydrogen, methyl or ethyl
radicals.
15. The composition of claim 13 wherein the aminophosphonic acid is
diethylenetriaminepentamethylenephosphonic acid or a physiologically
acceptable salt thereof.
16. The composition of claim 18 wherein the radionuclide is Yb-175.
17. The composition of claim 1 wherein the radionuclide is Yb-175 and the
aminophosphonic acid is at least one of nitrilotrimethylenephosphonic
acid, ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
diethylenetriaminepentamethylenephosphonic acid, or a physiologically
acceptable salt thereof.
18. The composition of claim 1 wherein the radionuclide is Ho-166 and the
aminophosphonic acid is at least one of
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
N-methylethylenediaminetrimethylenephosphonic acid,
N-isopropylethylenediaminetrimethylenephosphonic acid,
N-benzylethylenediaminetrimethylenephosphonic acid, or a physiologically
acceptable salt thereof.
19. The composition of claim 1 wherein the radionuclide is Lu-177 and the
aminophosphonic acid is at least one of
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid, or a
physiologically acceptable salt thereof.
20. The composition of claim 1 wherein the radionuclide is Gd-159 and the
aminophosphonic acid is at least one of
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid, or a
physiologically acceptable salt thereof.
21. A composition comprising 1) a complex which comprises at least one
radionuclide of Gd-159, Ho-166, Lu-177 or Yb-175 and at least one of an
aminophosphonic acid, or a physiologically acceptable salt thereof,
wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
diethylenetriaminepentamethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
nitrilotrimethylenephosphonic acid,
N-methylethylenediaminetrimethylenephosphonic acid,
N-isopropylethylenediaminetrimethylenephosphonic acid, or
N-benzylethylenediaminetrimethylenephosphonic acid, and 2) at least one of
the above aminophosphonic acids, or a physiologically acceptable salt
thereof, in excess of that required to make the complex, and wherein the
resulting composition is therapeutically effective.
22. The composition of claim 21 wherein a physiologically acceptable liquid
carrier is present.
23. The composition of claim 22 wherein the physiologically acceptable
liquid carrier is water and the resulting solution is adjusted to have a
pH of about 7 to about 8.
24. The composition of claim 21 wherein the radionuclide is Gd-159.
25. The composition of claim 24 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid, or a
physiologically acceptable salt thereof.
26. The composition of claim 21 wherein the radionuclide is Ho-166.
27. The composition of claim 26 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
N-methylethylenediaminetrimethylenephosphonic acid,
N-isopropylethylenediaminetrimethylenephosphonic acid,
N-benzylethylenediaminetrimethylenephosphonic acid, or a physiologically
acceptable salt thereof.
28. The composition of claim 21 wherein the radionuclide is Lu-177.
29. The composition of claim 28 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid, or a
physiologically acceptable salt thereof.
30. The composition of claim 21 wherein the radionuclide is Yb-175.
31. The composition of claim 30 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
diethylenetriaminepentamethylenephosphonic acid,
nitrilotrimethylenephosphonic acid, or a physiologically acceptable salt
thereof.
32. A sterile composition suitable for administration to a mammal
comprising 1) a complex which comprises at least one radionuclide of
Gd-159, Ho-166, Lu-177 or Yb-175 and at least one of an aminophosphonic
acid, or a physiologically acceptable salt thereof, wherein the
aminophosphonic acid is ethylenediaminetetramethylenephosphonic acid,
diethylenetriaminepentamethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
nitrilotrimethylenephosphonic acid,
N-methylethylenediaminetrimethylenephosphonic acid,
N-isopropylethylenediaminetrimethylenephosphonic acid, or
N-benzylethylenediaminetrimethylenephosphonic acid, and 2) at least one of
the above aminophosphonic acids, or a physiologically acceptable salt
thereof, in excess of that required to make the complex, and wherein the
resulting composition is therapeutically effective and wherein the
radionuclide in dosage form is present in an amount containing at least
0.02 mCi per kilogram of body weight of said mammal.
33. The composition of claim 32 wherein a physiologically acceptable liquid
carrier is present.
34. The composition of claim 33 wherein the physiologically acceptable
liquid carrier is water and the resulting solution is adjusted to have a
pH of about 7 to 8.
35. The composition of claim 32 wherein the radionuclide is Gd-159.
36. The composition of claim 35 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid, or a
physiologically acceptable salt thereof.
37. The composition of claim 32 wherein the radionuclide is Ho-166.
38. The composition of claim 37 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
N-methylethylenediaminetrimethylenephosphonic acid,
N-isopropylethylenediaminetrimethylenephosphonic acid,
N-benzylethylenediaminetrimethylenephosphonic acid, or a physiologically
acceptable salt thereof.
39. The composition of claim 32 wherein the radionuclide is Lu-177.
40. The composition of claim 39 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid, or a
physiologically acceptable salt thereof.
41. The composition of claim 32 wherein the radionuclide is Yb-175.
42. The composition of claim 41 wherein the aminophosphonic acid is
ethylenediaminetetramethylenephosphonic acid,
hydroxyethylethylenediaminetrimethylenephosphonic acid,
diethylenetriaminepentamethylenephosphonic acid,
nitrilotrimethylenephosphonic acid, or a physiologically acceptable salt
thereof.
Description
BACKGROUND OF THE INVENTION
The development of bone metastases is a common and often catastrophic event
for a cancer patient. The pain, pathological fractures, frequent
neurological deficits and forced immobility caused by these metastatic
lesions significantly decrease the quality of life for the cancer patient.
The number of patients that contract metastatic disease is large since
nearly 50% of all patients who contract breast, lung or prostate carcinoma
will eventually develop bone metastases. Bone metastases are also seen in
patients with carcinoma of the kidney, thyroid, bladder, cervix and other
tumors, but collectively, these represent less than 20% of patients who
develop bone metastases. Metastatic bone cancer is rarely life threatening
and occasionally patients live for years following the discovery of the
bone lesions. Initially, treatment goals center on relieving pain, thus
reducing requirements for narcotic medication and increasing ambulation.
Clearly, it is hoped that some of the cancers can be cured.
The use of radionuclides for treatment of cancer metastatic to the bone
dates back to the early 1950's. It has been proposed to inject a
radioactive particle-emitting nuclide in a suitable form for the treatment
of calcific lesions. It is desirable that such nuclides be concentrated in
the area of the bone lesion with minimal amounts reaching the soft tissue
and normal bone. Radioactive phosphorus (P-32 and P-33) compounds have
been proposed, but the nuclear and biolocalization properties limit the
use of these compounds. (Kaplan, E., et al, Journal of Nuclear Medicine,
Vol. 1, No. 1, page 1, 1960); (U.S. Pat. No. 3,965,254).
Another attempt to treat bone cancer has been made using phosphorus
compounds containing a boron residue. The compounds were injected into the
body (intravenously) and accumulated in the skeletal system. The treatment
area was then irradiated with neutrons in order to activate the boron and
give a therapeutic radiation dose. (U.S. Pat. No. 4,399,817).
In the above mentioned procedures, it is not possible to give therapeutic
doses to the tumor without substantial damage to normal tissues. In many
cases, especially for metastatic bone lesions, the tumor has spread
throughout the skeletal system and amputation or external beam irradiation
is not practical. (Seminars in Nuclear Medicine, Vol. IX, No. 2, April,
1979).
The use of Re-186 complexed with a diphosphonate has also been proposed.
(Mathieu, L. et al, Int. J. Applied Rad. & Isotopes, Vol. 30, pp. 725-727,
1979; Weinenger, J., Ketring, A. R., et al, Journal of Nuclear Medicine,
Vol. 24, No. 5, P125, 1983). However, the preparation and purification
needed for this complex limits its utility and wide application.
Strontium-89 has also been proposed for patients with metastatic bone
lesions. However, the long half-life (50.4 days), high blood levels and
low lesion to normal bone ratios limit the utility. (Firusian, N., Mellin,
P., Schmidt, C. G., The Journal of Urology, Vol. 116, page 764, 1976;
Schmidt, C. G., Firusian, N., Int. J. Clin. Pharmacol., 93:199-205, 1974).
A palliative treatment of bone metastases has been reported which employed
I-131 labelled .alpha.-amino-(3-iodo-4-hydroxybenzylidene)diphosphonate
(Eisenhut, M., Journal of Nuclear Medicine, Vol. 25, No. 12, pp.
1356-1361, 1984). The use of radioiodine as a therapeutic radionuclide is
less than desirable due to the well known tendency of iodine to localize
in the thyroid. Eisenhut lists iodide as one of the possible metabolites
of this compound.
SUMMARY OF THE INVENTION
Certain particle-emitting radionuclides, e.g. Samarium-153, have been
complexed with certain organic aminoalkylenephosphonic acids, i.e.
aminophosphonic acids in which the nitrogen and phosphorus atoms are
interconnected by an alkylene or substituted alkylene group, for example
ethylenediaminetetramethylenephosphonic acid, and physiologically
acceptable salts thereof. Certain compositions containing these complexes
have been found useful for therapy of calcific tumors in animals. The
administration of the therapeutic compositions has been palliative to the
animal, for example by alleviating pain and/or inhibiting tumor growth
and/or causing regression of tumors and/or destroying the tumors.
As used herein, the term "animals" includes humans; the term "calcific
tumors" includes primary tumors, where the skeletal system is the first
site of involvement, invasive tumors where the primary tumor invades the
skeletal system or other tissue tumors which calcify, and metastatic bone
cancer where the neoplasm spreads from other primary sites, e.g. prostate
and breast, into the skeletal system.
DETAILED DESCRIPTION OF THE INVENTION
The compositions of this invention are used for the therapeutic treatment
of calcific tumors in animals. These compositions contain certain
radionuclides complexed with certain aminophosphonic acids, or
physiologically acceptable salts thereof. As will be more fully discussed
later, the properties of the radionuclide, of the aminophosphonic acid and
of the complex formed therefrom are important considerations in
determining the effectiveness of any particular composition employed for
such treatment.
Particle-emitting radionuclides employed in the compositions of the
invention are capable of delivering a high enough localized ionization
density to alleviate pain and/or inhibit tumor growth and/or cause
regression of tumors, and/or destroy the tumor and are capable of forming
complexes with the aminophosphonate ligands described herein. The
radionuclides found to be useful in the practice of the invention are
Samarium-153 (Sm-153), Holmium-166 (Ho-166), Ytterbium-175 (Yb-175),
Lutetium-177 (Lu-177), and Gadolinium-159 (Gd-159).
The organic aminophosphonic acids which have been found useful in the
compositions of this invention are organic amine or substituted organic
amine compounds wherein the amine nitrogen and the phosphorus of the
phosphonic acid moiety are interconnected by an alkylene or substituted
alkylene group having the formula
##STR1##
wherein X and Y are independently selected from hydrogen, hydroxyl,
carboxyl, phosphonic, and hydrocarbon radicals having from 1-8 carbon
atoms and physiologically acceptable salts of the acid radicals and n is
1-3 with the proviso that when n>1 each X and Y may be the same as or
different from the X and Y of any other carbon atom.
The following structural formulas represent some of the ligands which can
be used in the compositions of this invention:
##STR2##
wherein substituents A, B, C, D, E and F are independently selected from
hydrogen, methyl, ethyl, isopropyl, benzyl,
##STR3##
and physiologically acceptable salts of the acid radicals wherein X, Y,
and n are as previously defined, X' and Y' are independently hydrogen,
methyl or ethyl radicals, n' is 2 or 3 and m and m' each is independently
0-10, with the proviso that at least one of said nitrogen substituents is
a phosphorus-containing group as previously defined herein, and wherein R
is a hydrocarbon residue which can be a linear, branched, cyclic,
heterocyclic, substituted heterocyclic, or a fused ring-type structure;
with the further proviso that when m or m'>1 the E and F substituents may
be the same as or different from any other substituent of any other
nitrogen atom and each R can be the same as or different from any other R.
In addition, cyclic amines containing the above mentioned substituents,
provided that at least one is a phosphorous-containing group as previously
defined, are useful in the compositions of the invention.
Some specific, but non-limiting, examples of ligands which are included by
the above structures are ethylenediaminetetramethylenephosphonic acid
(EDTMP), diethylenetriaminepentamethylenephosphonic acid (DTPMP),
hydroxyethylethylenediaminetrimethylenephosphonic acid (HEEDTMP),
nitrilotrimethylenephosphonic acid (NTMP),
tris(2-aminoethyl)aminehexamethylenephosphonic acid (TTHMP),
1-carboxyethylenediaminetetramethylenephosphonic acid (CEDTMP)
bis(aminoethylpiperazine)tetramethylenephosphonic acid (AEPTMP),
N-methylethylenediaminetrimethylenephosphonic acid (MEDTMP),
N-isopropylethylenediaminetrimethylenephosphonic acid (IEDTMP) and
N-benzylethylenediaminetrimethylenephosphonic acid (BzEDTMP).
For the purpose of the present invention, the complexes described herein
and physiologically acceptable salts thereof are considered equivalent in
the therapeutically effective compositions. Physiologically acceptable
salts refer to the acid addition salts of those bases which will form a
salt with at least one acid group of the ligand or ligands employed and
which will not cause a significant adverse physiological effect when
administered to an animal at dosages consistent with good pharmacological
practice. Suitable bases include, for example, the alkali metal and
alkaline earth metal hydroxides, carbonates, and bicarbonates such as
sodium hydroxide, potassium hydroxide, calcium hydroxide, potassium
carbonate, sodium bicarbonate, magnesium carbonate and the like, ammonia,
primary, secondary and tertiary amines and the like. Physiologically
acceptable salts may be prepared by treating the acid with an appropriate
base.
Complexes employed in the compositions of the present invention must fit
certain criteria insofar as possible. It should be recognized that there
are many ligands, or complexing agents, which are included by our
definition of the aminophosphonic acids in which the nitrogen of the amine
and the phosphorus of the phosphonic acid group are interconnected by an
alkylene group. Many may also contain other functional groups as
substituents for some, but not all, of the amine hydrogens of the ligand.
It should also be recognized that the properties of the particular
radionuclide are important. The disadvantage of any one property may be
overcome by the superiority of one or more of the properties of either
ligand or radionuclide and their combination as the complex must be
considered in toto.
The following is a discussion of the criteria which must be considered in
choosing any particular therapeutically effective combination of
radionuclide and ligand. Certain combinations, due to one or more
undesirable properties, may not be therapeutically useful or effective,
e.g. too much radioactivity localizing in non-osseous tissue.
The radionuclide must be delivered preferentially to the bone rather than
to soft tissue. Most particularly, uptake in either liver or bone marrow
is undesirable. Another important criterion is the ratio of the amount of
radionuclide taken up by the cancerous bone to that by normal bone. High
ratios are preferred. Also, the radionuclide should be cleared rapidly
from non-osseous tissue to avoid unnecessary damage to such tissues, e.g.,
it should clear rapidly from the blood.
For the purpose of convenience the abbreviations given hereinbefore will be
used hereinafter to denote the respective radionuclides and
aminophosphonic acids.
Preferred embodiments of the present invention are therapeutically
effective or useful compositions containing complexes of at least one
radionuclide selected from Gd-159, Ho-166, Lu-177, Sm-153 and Yb-175 with
at least one ligand selected from EDTMP, DTPMP, HEEDTMP, NTMP, TTHMP,
CEDTMP, AEPTMP, MEDTMP, IEDTMP, BzEDTMP, or a physiologically acceptable
salt(s) thereof. Preferably a physiologically acceptable liquid carrier is
also present with the complexes of the present invention.
Not all compositions containing radionuclides and ligands are
therapeutically useful. Thus, for example, a composition containing Sm-153
and monoethanolaminedimethylenephosphonic acid was unacceptable because a
significant portion of the radioactivity localized in the liver.
Combinations of the various above noted radionuclides can be administered
for the therapeutic treatment of calcific tumors. The combinations can be
complexed as herein described by complexing them simultaneously, mixing
two separately complexed radionuclides, or administering two different
complexed radionuclides sequentially. It may be possible to achieve the
same beneficial results of high delivery of the radionuclide to the area
of the tumor, but with little soft tissue damage, by administering the
ligand and the radionuclide in a manner which allows formation of the
radionuclide-chelant complex in situ such as by simultaneous or near
simultaneous administration of the radionuclide and an appropriate amount
of ligand or by the administration of ligand and a radionuclide complexed
with a weaker ligand, i.e., one which undergoes ligand exchange with the
ligands of this invention, such that the desired radionuclide-chelant
complex is formed via ligand exchange in situ. The composition may be
administered as a single dose or as multiple doses over a longer period of
time.
The aminophosphonic acids can be prepared by a number of known synthetic
techniques. Of particular importance is the reaction of a compound
containing at least one reactive amine hydrogen with a carbonyl compound
(aldehyde or ketone) and phosphorous acid or derivative thereof.
Amine precursors employed in making the aminophosphonic acids employed in
the present invention are commercially available products or may be
prepared readily by methods known to those skilled in the art of organic
synthesis.
Methods for carboxyalkylating to obtain the amine derivatives containing
one or more carboxyalkyl groups are well known (U.S. Pat. No. 3,726,912)
as are the methods which give alkyl phosphonic and hydroxyalkyl (U.S. Pat.
No. 3,398,198) substituents on the amine nitrogens.
Radionuclides can be produced in several ways. In a nuclear reactor, a
nuclide is bombarded with neutrons to obtain a radionuclide, e.g.
##STR4##
Another method of obtaining radionuclides is by bombarding nuclides with
linear accelerator or cyclotron-produced particles. Yet another way of
obtaining radionuclide3 is to isolate them from fission product mixtures.
The method of obtaining the radionuclide is not critical to the present
invention.
To irradiate Sm.sub.2 O.sub.3 for production of Sm-153, the desired amount
of target was first weighed into a quartz vial, the vial was flame sealed
under vacuum and welded into an aluminum can. The can was irradiated for
the desired length of time, cooled for several hours and opened remotely
in a hot cell. The quartz vial was removed and transferred to a glove box,
crushed into a glass vial which was then sealed with a rubber septum and
an aluminum crimp cap. One milliliter of 1-4M HCl was then added to the
vial via syringe to dissolve the Sm.sub.2 O.sub.3. Once dissolved, the
solution was diluted to the appropriate volume by addition of water. The
solution was removed from the original dissolution vial which contains
chards of the crushed quartz vial and transferred via syringe to a clean
glass serum vial. This solution was then used for complex preparation.
Similar procedures were used to prepare Lu-177, Yb-175, Gd-159, and
Ho-166.
When aqueous solutions of metal ions are mixed with solutions containing
complexing agents, such as those described in this invention, a complex
between the metal ion and the ligand can be formed as shown by the
equation below.
M+L.revreaction.M.multidot.L
The reaction is believed to be an equilibrium such that the concentrations
of metal (M) and complexing agent, or ligand (L), can affect the
concentration of species present in solution. Competing side reactions,
such as metal hydroxide formation, can also occur in aqueous solution,
thus
xM+yOH.sup.- .fwdarw.M.sub.x (OH)y.
The OH.sup.- concentration in solution, which is related to pH is,
therefore, an important parameter to be considered. If the pH is too high,
the metal tends to form metal hydroxides rather than complexes. The
complexing agents may also be adversely affected by low pH. Complexation
may require the loss of proton(s); therefore at low pH, conditions may not
be favorable for complexation to occur. Consideration must be given to the
solubility characteristics of the ligand, radionuclide, and complex.
Although not limited thereto, a pH in the range of from 5 to 11 is
preferred for complexation.
The metal and ligand may be combined under any conditions which allow the
two to form a complex. Generally, mixing in water at a controlled pH (the
choice of pH is dependent upon the choice of ligand and metal) is all that
is required.
The ratio of ligand to metal is a result of two competing considerations.
As indicated above, the ligand and metal are believed to be in equilibrium
with the complex. As appreciated by one skilled in the art of
radiochemistry, only a portion of the metal which is irradiated will
become radioactive. In the practice of the invention it is preferred that
an amount of the ligand be employed that is sufficient to insure that all
of the radionuclide present is complexed since any uncomplexed
radionuclide may localize in soft tissue. To insure complexation of the
radionuclide it is preferred that the amount of ligand used be in excess
of the total amount of metal present, i.e. radioactive metal plus
non-radioactive metal plus any other metals present that can complex with
the ligand. Thus, in the practice of the invention it is desirable to
employ the complexed nuclide in the presence of an excess of ligand. Such
excess should provide an amount sufficient to inhibit significant uptake
of the radionuclide by soft tissue. The excess ligand may be the same as
or different from that used to complex the radionuclide. However, too much
free ligand may have adverse effects, e.g. it may be toxic to the patient
or result in less favorable biolocalization of the radionuclide.
Most of the complexes employed in this invention were prepared as follows:
the desired amount of ligand was placed in a vial and dissolved by
addition of water. At some higher ligand concentrations, it was necessary
to add base in order to completely dissolve the ligand. Heating was also
found to be useful for dissolving the ligands. The appropriate amount of
the samarium or other radionuclides in the stock solution described above
was then added to the ligand solution. The pH of the resulting solution
was then raised to the appropriate level (usually 7-8) by addition of
NAOH. It is also possible to prepare the complexes of the present
invention in physiologically acceptable liquid carriers other than water.
Useful solvents include aqueous alcohols, glycols, and phosphonate or
carbonate esters.
The invention described herein provides a means of delivering a therapeutic
amount of radioactivity to calcific tumors. However, it may also be
desirable to administer a "sub-therapeutic" amount to determine the fate
of the radionuclide using a scintillation camera prior to administering a
therapeutic dose. Therapeutic doses will be administered in sufficient
amounts to alleviate pain and/or inhibit tumor growth and/or cause
regression of tumors and/or kill the tumor. Amounts of radionuclide needed
to provide the desired therapeutic dose will be determined experimentally
and optimized for each particular composition. The amount of radioactivity
required to deliver a therapeutic dose will vary with the individual
composition employed. The composition to be administered may be given in a
single treatment or fractionated into several portions and administered at
different times. Administering the composition in fractionated doses may
make it possible to minimize damage to non-target tissue. Such multiple
dose administration may be more effective.
The compositions of the present invention may be admisistered as a sterile
composition and may be used in conjunction with other active agents and/or
ingredients that enhance the therapeutic effectiveness of the compositions
and/or facilitate easier administration of the compositions.
Studies to determine the qualitative biodistribution of the various
radionuclides were conducted by injecting the compositions into rats and
obtaining the gamma ray images of the entire animal at various times up to
two hours after injection.
Quantitative biodistributions were obtained by injecting 50-100 microliters
of the composition into the tail vein of unanesthetized male Sprague
Dawley rats. The rats were then placed in cages lined with absorbent paper
in order to collect all urine excreted prior to sacrifice. After a given
period of time, the rats were sacrificed by cervical dislocation and the
various tissues dissected. The samples were then rinsed with saline,
blotted dry on absorbent paper and weighed. The radioactivity in the
samples was measured with a NaI scintillation counter.
Biolocalization studies were also performed in rabbits. The rabbits were
injected with 100-250 microliters of the composition into the marginal ear
vein. In studies where blood clearance was measured, blood samples were
taken through a heparinized cannula placed in the marginal vein of the ear
not used for injection. Three hours after injection, a blood sample was
taken from each rabbit by cardiac puncture and each animal was sacrificed
by injection of a commercial euthanasia solution. After sacrifice, images
were obtained by placing the carcass directly on the face of a large
field-of-view scintillation camera.
Uptake of the radionuclide in the lesion compared to uptake in normal bone,
or lesion/normal bone ratios, is a particularly important parameter for
determining the suitability of any composition for therapy. To determine
the lesion to normal bone ratios, a modified drill hole method
(Subramanian, G. et al., 19th Intl. Annual Meetings of S. N. M., Bern,
Switzerland, Sep. 8-11, 1981) was used. Thus, two holes were drilled into
the surface of the tibia of a rabbit in order to damage the bone. Seven to
ten days later, the animal was injected with the radionuclide composition.
After three hours, the rabbit was anesthetized and imaged using an Anger
camera and pinhole collimator.
The compositions of the present invention were also shown to relieve bone
pain. The compositions go preferentially to bone and are effective as are
the compositions for calcific tumor treatment.
The following examples show representative preparations of aminophosphonic
acid ligands and complexes of the ligands with the radionuclides and the
use of these compositions.
GENERAL EXPERIMENTAL
Whenever in the following examples and tables data is presented on the
administered dose of radioactivity, the numbers given represent the
percentage of the administered dose which localized in the indicated
tissue. The ratios of radioactivity observed in bone relative to blood and
muscle were calculated based on the percent dose per gram in the bone and
in the blood and muscle. Whenever the ratios of radioactivity found in
bone to that found in nonosseous tissue is given, the calculation was made
as indicated above on a percent dose per gram basis.
EXAMPLE 1
Into a suitable reaction vessel equipped with a thermometer, magnetic
stirring bar, dropping funnel, and an atmosphere of nitrogen were charged
phosphorous acid (94.5 g) and degassed water (100 ml). Dissolution of the
phosphorous acid was achieved by stirring and then concentrated
hydrochloric acid (112 ml) was added. The dropping funnel was charged with
ethylenediamine (15 g) and adjusted to allow dropwise addition of the
amine to the acidic solution. When addition was complete a heating mantle
was installed and the solution refluxed for one hour. At the end of this
time the dropping funnel was charged with formaldehyde (85 g of a 37%
aqueous solution) which was added dropwise over a two hour period with
continued heating to maintain reflux during the addition. After all the
formaldehyde was added, the reaction mixture was stirred under reflux for
an additional two hours, then allowed to cool slowly overnight during
which time the product precipitated. Vacuum filtration followed by cold
water washing yielded ethylenediaminetetramethylenephosphonic acid
(EDTMP).
EXAMPLE 2
A quantity of 25 to 35 milligrams of EDTMP was weighed into a vial and
dissolved using 0.75 ml of distilled water. To this, 0.25 ml of Sm-153
(.about.10 mCi) in dilute HCl was added. The pH of the resulting solution
was then adjusted to 10 by addition of NAOH. The resulting solution was
heated to between 60.degree.-70.degree. C. for 30 minutes in a water bath.
The pH of the solution was then adjusted to 7-8 by addition of HCl.
Laboratory rats were injected with the above complex (50-100 .mu.l) via the
tail vein. After 2 hours, the animals were sacrificed by cervical
dislocation and organs and tissues removed. Samples were counted with a
NaI counter to determine the biolocalization of the radionuclide. It was
found that a significant amount (55-65%) of the radioactivity was
concentrated in the skeletal system with very little soft tissue uptake.
Most of the radioactivity not found in the skeleton was cleared through
the kidneys into the urine. Scintillation scans of animals treated in the
same manner showed the radioactivity concentrating in the skeletal system.
The lesion to normal bone ratio was approximately equal to that of
Tc-99m-MDP (MD? refers to methylene diphosphonate), a commercially
available diagnostic bone agent.
EXAMPLE 3
Into a suitable reaction vessel equipped with a thermometer, magnetic
stirring bar, dropping funnel, and an atmosphere of nitrogen were charged
phosphorous acid (94.5 g) and degassed water (100 ml). Dissolution of the
phosphorous acid was achieved by stirring and concentrated hydrochloric
acid (112 ml) was added. The dropping funnel was charged with
diethylene-triamine (20.6 g) and adjusted to allow dropwise addition of
the amine to the acidic solution. When addition was complete a heating
mantle was installed and the solution refluxed for one hour. At the end of
this time the dropping funnel was charged with formaldehyde (85 g of a 37%
aqueous solution) which was added dropwise over a two hour period with
continued heating to maintain reflux during the addition. After all the
formaldehyde was added, the reaction mixture was stirred under reflux for
an additional two hours, then allowed to cool.
Diethylenetriaminepentamethylenephosphonic acid (DTPMP) was isolated from
the reaction mixture.
EXAMPLE 4
A quantity of 20 to 30 milligrams of DTPMP was weighed into a vial and
dissolved using 0.75 ml of distilled water. To this, 0.25 ml of Sm-153
(.about.10 mCi) in dilute HCl was added. The pH of the resulting solution
was then adjusted to 10 by addition of NAOH. The solution was then heated
to between 60.degree.-70.degree. C. for 30 minutes in a water bath. The pH
of the solution was then adjusted to 7-8 by addition of HCl. This
composition was then injected into rats and the biolocalization of Sm-153
was determined.
EXAMPLE 5
Into a suitable reaction vessel equipped with a thermometer, magnetic
stirring bar, dropping funnel, and an atmosphere of nitrogen were charged
phosphorous acid (94.5 g) and degassed water (100 ml). Dissolution of the
phosphorous acid was achieved by stirring and concentrated hydrochloric
acid (112 ml) was added. The dropping funnel was charged with
N-hydroxyethylethylenediamine (34.6 g) and adjusted to allow dropwise
addition of the amine to the acidic solution. When addition was complete a
heating mantle was installed and the solution refluxed for one hour. At
the end of this time the dropping funnel was charged with formaldehyde (85
g of a 37% aqueous solution) which was added dropwise over a two hour
period with continued heating to maintain reflux during the addition.
After all the formaldehyde was added, the reaction mixture was stirred
under reflux for an additional two hours, then allowed to cool.
Hydroxyethylethylenediaminetrimethylenephosphonic acid (HEEDTMP) was
isolated from the reaction mixture.
EXAMPLE 6
A quantity of 30 to 40 milligrams of HEEDTMP was weighed into a vial and
dissolved using 0.75 ml of distilled water. To this, 0.25 ml of Sm-153
(.about.10 mCi) in dilute HCl was added. The pH of the resulting solution
was then adjusted to 10 by addition of NAOH. The solution was then heated
between 60.degree.-70.degree. C. for 30 minutes in a water bath. The pH of
the solution was then adjusted to 7-8 by addition of HCl. This composition
was then injected into rats and the biolocalization of Sm-153 was
determined.
EXAMPLE 7
Into a suitable reaction vessel equipped with a thermometer, magnetic
stirring bar, dropping funnel, and an atmosphere of nitrogen were charged
phosphorous acid (57.7 g) and degassed water (50 ml). Dissolution of the
phosphorous acid was achieved by stirring and concentrated hydrochloric
acid (50 ml) was added. The dropping funnel was charged with tris
(2-aminoethyl)amine (13.7 g) and adjusted to allow dropwise addition of
the amine to the acidic solution. When addition was complete a heating
mantle was installed and the solution refluxed for one hour. At the end of
this time the dropping funnel was charged with formaldehyde (51 g of a 37%
aqueous solution) which was added dropwise over a two hour period with
continued heating to maintain reflux during the addition. After all the
formaldehyde was added, the reaction mixture was stirred under reflux for
an additional two hours, then allowed to cool.
Tris(2-aminoethyl)aminehexamethylenephosphonic acid (TTHMP) was isolated.
EXAMPLE 8
A quantity of 48 to 53 milligrams of TTHMP was weighed into a vial and
dissolved using 0.75 ml of distilled water. To this solution, 0.25 ml of
Sm-153 (.about.10 mCi) in dilute HCl was added. The pH of the resulting
solution was adjusted to 10 by addition of NAOH. The solution was then
heated between 60.degree.-70.degree. C. for 30 minutes in a water bath.
The pH of the solution was then adjusted to 7-8 by addition of HCl. This
composition was then injected into rats and the biolocalization of Sm-153
was determined.
The data obtained with respect to the two hour biolocalization of Sm-153 in
rats for the compositions of Examples 2, 4, 6 and 8 is shown in Table I.
TABLE I
______________________________________
Example Nos.
% Dose in 2.sup.a
4.sup.a 6.sup.a
8.sup.a
______________________________________
Skeleton 58 30 57 28
Blood 0.032 0.16 0.035
0.25
Liver 0.25 0.27 0.45 0.18
Urine 49 74 50 65
Bone/Blood 1800 224 1300 80
Bone/Muscle 1500 220 1300 410
______________________________________
.sup.a = represent the average of the results of 5 rats
EXAMPLE 9
A quantity of 35 to 45 milligrams of EDTMP was weighed into a vial and
dissolved using 0.75 ml of distilled water. To this solution, 0.25 ml of
Yb-175 in dilute HCl was added. The pH of the resulting solution was
adjusted to 10 by addition of NAOH. The solution was then heated between
60.degree.-700.degree. C. for 30 minutes in a water bath. The pH of the
solution was then adjusted to 7-8 by addition of HCl.
EXAMPLE 10
A quantity of 55 to 60 milligrams of DTPMP was weighed into a vial and
dissolved using 0.75 ml of distilled water. To this solution, 0.25 ml of
Yb-175 in dilute HCl was added. The pH of the resulting solution was
adjusted to 10 by addition of NAOH. The solution was then heated between
60.degree.-70.degree. C. for 30 minutes in a water bath. The pH of the
solution was then adjusted to 7-8 by addition of HCl.
EXAMPLE 11
A quantity of 50 to 55 milligrams of HEEDTMP was weighed into a vial and
dissolved with 0.75 ml of distilled water. To this solution, 0.25 ml of
Yb-175 in dilute HCl was added. The pH of the resulting solution was
adjusted to 10 by addition of NAOH. The solution was then heated between
60.degree.-70.degree. C. for 30 minutes in a water bath. The pH of the
solution was then adjusted to 7-8 by addition of HCl.
EXAMPLE 12
Into a suitable reaction vessel equipped with a thermometer, magnetic
stirring bar, dropping funnel, and an atmosphere of nitrogen were charged
phosphorous acid (94.5 g) and degassed water (100 ml). Dissolution of the
phosphorous acid was achieved by stirring and concentrated hydrochloric
acid (112 ml) was added. The dropping funnel was charged with ammonium
chloride (17.2 g in an aqueous solution) and adjusted to allow dropwise
addition of the ammonium chloride to the acidic solution. When addition
was complete a heating mantle was installed and the solution refluxed for
one hour. At the end of this time the dropping funnel was charged with
formaldehyde (85 g of a 37% aqueous solution) which was added dropwise
over a two hour period with continued heating to maintain reflux during
the addition. After all the formaldehyde was added, the reaction mixture
was stirred under reflux for an additional two hours, then allowed to
cool, yielding nitrilotrimethylenephosphonic acid (NTMP).
EXAMPLE 13
A quantity of 50 to 55 milligrams of NTMP was weighed into a vial and
dissolved with 0.75 ml of distilled water. To this solution, 0.25 ml of
Yb-175 in dilute HCl wa3 added. The pH of the resulting solution was then
adjusted to 10 by addition of NAOH. The solution was then heated between
60.degree.-700.degree. C. for 30 minutes. The pH of the solution was then
adjusted to 7-8 by addition of HCl.
The compositions of Examples 9, 10, 11 and 13 were each injected into rats
and the two hour biolocalization of Yb-175 for each of these compositions
was determined; the data obtained is shown in Table II.
TABLE II
______________________________________
Example Nos.
% Dose in 9.sup.a
10.sup.a 11.sup.a
13.sup.a
______________________________________
Skeleton 50 25 56 63
Blood 0.074 0.116 0.231
0.204
Liver 0.138 0.112 0.214
0.236
Bone/Blood 562 179 190 256
Bone/Muscle 619 366 572 876
______________________________________
.sup.a = represent the average of the results of 5 rats
EXAMPLE 14
A quantity of 50 to 55 milligrams of EDTMP was weighed into a vial and
dissolved with 0.75 ml of distilled water. To this solution, 0.25 ml of
Lu-177 in dilute HCl was added. The pH of the resulting solution was
adjusted to 10 by addition of NAOH. The solution was then heated between
60.degree.-70.degree. C. for 30 minutes. The pH of the solution was then
adjusted to 7-8 by addition of HCl.
EXAMPLE 15
A quantity of 55 to 60 milligrams of HEEDTMP was weighed into a vial and
dissolved with 0.75 ml of distilled water. To this, 0.25 ml of Lu-177 in
dilute HCl was added. The pH of the resulting solution was adjusted to 10
by addition of NAOH. The solution was then heated between
60.degree.-70.degree. C. for 30 minutes. The pH of the solution was then
adjusted to 7-8 by addition of HCl.
EXAMPLE 16
A quantity of 25 to 35 milligrams of EDTMP was weighed into a vial and
dissolved with 0.75 ml of distilled water. To this solution 0.25 ml of
Ho-166 in dilute HCl was added. The pH of the resulting solution was
adjusted to between 7-8.
The compositions of Examples 14, 15, and 16 were each injected into rats
and the two hour biolocalization of the radionuclide for each of the
compositions was determined; the data obtained is shown in Table III.
TABLE III
______________________________________
Example Nos.
% Dose in 14.sup.a 15.sup.a
16.sup.b
______________________________________
Skeleton 51 58 48
Blood 0.10 0.17 0.03
Liver 0.48 1.9 0.05
Kidneys 0.28 0.30 0.26
Muscle 0.44 0.56 0.10
Bone/Blood 500 370 1114
Bone/Muscle 1380 570 2292
______________________________________
.sup.a = represent the average of the results of 5 rats
.sup.b = represent the average of the results of 3 rats
EXAMPLE 17
Bis(aminoethyl)piperazinetetramethylenephosphonic acid (AEPTMP) was
prepared from bis(aminoethyl)piperazine in a manner similar to Example 1.
AEPTMP was complexed with Sm-153 and the biolocalization of the Sm-153
determined in rats.
EXAMPLE 18
1-Carboxyethylenediaminetetramethylenephosphonic acid (CEDTMP) was prepared
from 1-carboxyethylenediamine in a manner similar to Example 1. CEDTMP was
complexed with Sm-153 and the biolocalization of the SM-153 determined in
rats.
The data obtained with respect to the two hour biolocalization of Sm-153 in
rats for the compositions of Examples 17 and 18 is shown in Table IV.
TABLE IV
______________________________________
Example No.
% Dose in 17.sup.a
18.sup.b
______________________________________
Skeleton 48 57
Blood 0.24 0.27
Liver 2.1 2.2
Muscle 0.65 1.4
Bone/Blood 150 217
Bone/Muscle 364 376
______________________________________
.sup.a = represent the average of the results of 4 rats
.sup.b = represent the average of the results of 2 rats
EXAMPLE 19
A quantity of 48 to 53 milligrams of EDTMP was weighed into a vial and
dissolved with 0.75 ml of distilled water. To this solution, 0.25 ml of
Gd-159 in dilute HCl was added. The pH of the resulting solution was
adjusted to 10 by addition of NaOH. The solution was then heated between
60.degree.-70.degree. C. in a water bath. The pH of the solution was then
adjusted to 7-8 by addition of HCl.
EXAMPLE 20
A quantity of 55 to 60 milligrams of HEEDTMP was weighed into a vial and
dissolved with 0.75 ml of distilled water. To this solution, 0.25 ml of
Gd-159 in dilute HCl was added. The pH of the resulting solution was
adjusted to 10 by addition of NaOH. The solution was then heated between
60.degree.-70.degree. C. in a water bath. The pH of the solution was then
adjusted to 7-8 by addition of HCl.
The data obtained with respect to the two hour biolocalization of Gd-159 in
rats for the compositions of Examples 19 and 20 is shown in Table V.
TABLE V
______________________________________
Example Example
% Dose in No. 19.sup.a
No. 20.sup.a
______________________________________
Skeleton 57 60
Blood 0.15 0.14
Liver 0.25 0.57
Kidneys 0.33 0.58
Muscle 0.56 0.76
Bone/Blood 305 335
Bone/Muscle 577 548
______________________________________
.sup.a = the average of the results of five rats
EXAMPLE 21
A series of rats was injected with compositions containing Sm-153 EDTMP
(compositions prepared as in Example 2) and sacrified in groups of five at
various intervals. The biolocalization data, along with the calculated
bone to blood and bone to muscle ratios, is summarized in Table VI. The
data shows rapid uptake of the radioactivity in bone and rapid blood
clearance, as well as no significant clearance of the radioactivity from
the skeletal system throughout the course of the experiment.
TABLE VI
__________________________________________________________________________
Average Percent of Injected Dose at:
15 30 1 2 5 24 48 72
Organ Min.
Min.
Hour
Hours
Hours
Hours
Hours
Hours
__________________________________________________________________________
Skeleton
48 53 58
58 59 52 60 57
Blood 5.852
2.304
.532
.032 .008
.007 .006
.006
Liver .959
.526
.322
.252 .370
.349 .458
.492
Kidneys
1.745
.805
.466
.254 .364
.250 .286
.216
Urine 28 42 47
49 46 55 50 53
Bone/Muscle
30 70 300
1500 3600
2400 2800
3400
Bone/Blood
7 20 120
1800 4200
6700 8200
7800
__________________________________________________________________________
EXAMPLE 22
Compositions containing Sm-153-EDTMP (compositions prepared as in Example
2) were also tested in rabbits. Results of three hour biolocalization
testing (averaged for 5 rabbits) are summarized in Table VII.
TABLE VII
______________________________________
% Dose in
______________________________________
Skeleton 66
Blood 0.12
Liver 0.95
Urine 34
Bone Marrow
0.15
Bone/Blood
900
Bone/Muscle
1200
______________________________________
EXAMPLE 23
An English Setter with a tumor in his pelvis was injected with
approximately 19 mCi of a composition containing Sm-153-EDTMP. After 2
hours, the dog was imaged using a scintillation camera. The scintillation
scan showed high uptake of radioactivity in the area of the tumor and
looked very similar to an earlier scan performed using Tc-99m-MDP. The
lesion to normal bone ratio for Sm-153 was very similar to that for
To-99m. The scintillation scan of the dog five days after treatment was
similar to the scintillation scan obtained two hours post injection. Seven
days after the treatment the dog appeared to be in less pain as evidenced
by an increase in its mobility.
EXAMPLE 24
An Irish Setter with a tumor in the femur was injected with a composition
containing Sm-153-EDTMP. The leg was amputated and samples of the tumor
and normal bone were analyzed for Sm-153. The amount of Sm-153 in the
tumor on a per gram basis was 15-20 times that in non-lesionous bone from
the same leg.
EXAMPLE 25
An Irish setter with undifferentiated sarcoma metastatic to bone was
treated with 16 mCi (0.57 mCi/kg) of a composition containing
Sm-153-EDTMP. Scintillation scans showed high uptake of Sm-153 in the area
of the lesion and were very similar to scans obtained earlier using
Tc-99m-MDP. Seven days after treatment the dog appeared to be in less pain
as evidenced by an increase in mobility and a decrease in lameness. After
approximately 3 months, when signs of lameness began to reappear, the dog
was given a second treatment of 15 mCi (0.49 mCi/kg), of a composition
containing Sm-153-EDTMP. Once again, after seven days the dog showed a
general increase in mobility and decrease in lameness. X-rays taken five
months after the second treatment demonstrated significant regression of
bone lesions. Eight months after the second treatment the dog had no
recurrence of lameness, had gained weight and appeared healthy. Nine
months after the second treatment the dog died of kidney failure. Total
necropsy found no evidence of cancer.
EXAMPLE 26
A series of normal beagles was treated with various dose levels of a
composition containing Sm-153-EDTMP. White blood cell and platelet counts
were measured weekly to check for signs of bone marrow supression. Dogs
given a dose of 1.0 mCi Sm-153 per kilogram of body weight showed a slight
decrease in both platelets and white blood cells 2-3 weeks after
administration of the composition. White cells and platelets, returned to
normal levels 4-5 weeks after administration.
Dogs given a dose of 0.5 mCi of Sm-153 per kilogram of body weight showed a
smaller decrease in the levels of platelets and white blood cells. The few
animals that did fall below normal platelet and white blood cell levels
2-3 weeks after administration of the Sm-153-EDTMP composition, returned
to normal levels 4-5 weeks after administration.
EXAMPLE 27
Sterile pyrogen-free vials were prepared to contain 210 mg of EDTMP and 140
mg NAOH by freeze-drying a solution containing appropriate amounts of
EDTMP and NAOH. The vials were sealed under vacuum. To prepare one of the
compositons of the invention, 6 ml of a solution containing Sm-153
(approximately 3.times.10.sup.-4 M in Sm) in 0.1M HCl was injected into
the sealed vial through a septum. The resulting solution had a pH of 7-8.
Five humans with metastatic bone cancer (three with lung cancer and two
with prostate cancer as the primary site) were injected with a composition
containing 2 mCi of Sm-153-EDTMP. Blood samples were taken intermittently
to determine the fraction of the Sm-153 remaining in the blood. The
results shown in Table VIII show that the radioactivity cleared very
rapidly from the blood. There was no significant concentration of
radioactivity in red cells since all of the measured activity was in the
plasma fraction.
Urine was collected and counted to determine the percentage of the
radioactivity excreted as a function of time after injection. The results
in Table IX show that the fraction of radioactivity which cleared into the
urine cleared rapidly.
Scintigraphic images were obtained two hours post injection and were used
for comparison with images previously obtained using Tc-99m-HDP
(hydroxymethylenediphosphonate), a commercially available diagnostic bone
imaging agent. These images indicated that the Sm-153 localized in
skeletal tissue with no significant uptake and retention in non-osseous
tissues indicating a rapid and efficient clearing of Sm-153 from the blood
and all other non-osseous tissues.
In addition, there is no observable retention of radioactivity in the
kidneys, even in the two hour post injection images. This contrasts with
the two hour images with Tc-99m HDP in which the kidneys were visualized.
Vital signs were measured at several post injection times and were not
significantly different than preinjection values. No significant changes
in blood profiles (e.g., white blood cell count, red blood cell count or
platelet count) or in serum chemistry profiles were observed in
post-injection blood samples as compared to the preinjection sample
values.
Qualitative comparisons of images obtained using compositions containing
Tc-99m-HDP and compositions containing Sm-153-EDTMP demonstrate that all
cancerous lesions observable with Tc-99m-HDP are seen equally well with
Sm-153-EDTMP.
The uptake observed for each radionuclide in selected lesions compared to
its uptake in normal bone was determined using digitized images. The
quantitative comparisons of the radioactivity in lesions and in normal
bone show that compositions containing Sm-153-EDTMP and compositions
containing Tc-99m.-HDP result in localization of radioactivity in the
lesions to a similar extent.
The results obtained in the testing with humans show an overall
biolocalization profile, as well as a Sm-153 concentration in cancerous
lesions which was high, selective and similar to that observed in dogs.
TABLE VIII
______________________________________
Blood Clearance in Humans with Metastatic Bone Disease
Percent Injected Dose Remaining in Whole Blood
Patient 5 15 30 60 120 180 240
No. min. min. min. min. min. min. min.
______________________________________
1 36.34 27.62 18.69 11.63
6.23 3.74 2.28
2 27.98 17.61 15.18 9.07 5.51 3.75 2.68
3 22.25 14.25 9.93 6.04 3.41 2.03 1.31
4 22.33 19.09 16.64 10.08
5.35 3.55 1.86
5 49.24 25.91 18.41 10.84
5.35 3.32 2.29
Average 31.63 20.90 15.77 9.53 5.17 3.28 2.08
______________________________________
TABLE IX
______________________________________
Percent Excretion in Urine in Humans
with Metastatic Bone Disease
Patient 12-24 24 Hr
No. 1 Hr. 2 Hr. 4 Hr.
4-8 Hr.
8-12 Hr.
Hr. Total
______________________________________
1 26.09 10.71 8.84 11.69 1.67 1.63 60.63
2 32.28 2.84 19.30
12.03 2.17 1.29 60.91
3 25.55 4.81 8.29 3.18 1.72 0.87 44.42
4 11.37 9.42 9.87 12.94 2.24 0.85 46.69
5 35.36 7.96 8.47 5.29 0.98 0.73 58.79
Average
26.13 7.15 10.95
9.02 1.76 1.07 56.09
______________________________________
EXAMPLE 28
Into a suitable vessel equipped with a thermometer, condenser, and magnetic
stirring bar were charged phosphorous acid (56.8 g) and water (70 ml).
Dissolution of the phosphorous acid was by stirring and then concentrated
HCL (60 ml) was added. To the resulting solution was added 15.6 g of
N-methylethylenediamine (Aldrich Chemical Co., Milwaukee, Wis.) all at
once. A heating mantle was installed and the solution heated to reflux.
Paraformaldehyde (19.9 g) was added in small portions to the refluxing
solution over a 6.5 hour period after which the solution was allowed to
continue refluxing for an additional 3.5 hours. After cooling to room
temperature the solution was poured into methanol with vigorous Stirring
to produce a white semisolid. The semisolid was isolated by vacuum
filtration and air dried to give
N-methyl-ethylenediaminetrimethylenephosphonic acid (MEDTMP) as a white
solid, melting point 176.degree. C. (decomposed). This product showed the
expected two singlets in a 2:1 ratio in the decoupled phosphorous-31 NMR
spectrum run on a Bruker Ac-250 MHz NMR spectrometer, pH 2.5 85%
phosphoric acid as external standard.
.sup.31 P NMR (D.sub.2 O): .delta. 10.78 (s, 2P), 7.29 (s, 1P).
EXAMPLE 29
A 79.2 milligram portion of MEDTMP was weighed into a vial and dissolved in
1.5 ml of HoCl.sub.3 (2.6.times.10.sup.4 M in 0.046N HCL spiked with
Ho-166 isotope) by the addition of 50% NAOH. The final pH was 7.5 and the
percent of holmium complexed as determined by cation exchange
chromatography was greater than 98%.
EXAMPLE 30
Into a suitable reaction vessel equipped with a thermometer, condenser and
magnetic stirring bar were charged phosphorous acid (14.2 g) and water (25
ml). Dissolution of the phosphorous acid was achieved by stirring and then
concentrated HCl (15 ml) was added. To the resulting solution was added
5.21 g of N-isopropylethylenediamine (Aldrich Chemical Co., Milwaukee,
Wis.) all at once. A heating mantle was installed and the solution heated
to reflux. Paraformaldehyde (4.98 g) was added in small portions to the
refluxing solution over a 1.5 hour period after which the solution was
allowed to continue refluxing for an additional 3.5 hours. After cooling
to room temperature, the solution was concentrated under vacuum to low
volume and poured into absolute ethanol with vigorous stirring to produce
a white semisolid. The semisolid was isolated by vacuum filtration and
dried under vacuum to give 17.56 g (90.5% yield) of
N-isopropylethylenediaminetrimethylenephosphonic acid (IEDTMP), as a white
hygroscopic solid. The NMR spectra was obtained with a Bruker Ac-250 MHz
NMR spectometer, 85% phosphoric acid as external standard.
.sup.31 P (D.sub.2 O): .delta. 10.05 (s, 2P), 8.69 (s, 1P).
EXAMPLE 31
A 79.3 milligram portion of IEDTMP was dissolved in 1.5 mL of HoCl.sub.3
(2.6.times.10.sup.-4 M in 0.046N HCl spiked with Ho-166 isotope) by the
addition of 50% NAOH. The final PH was 7.5 and the percent of holmium
complexed was determined by cation exchange chromatography to be greater
than 97%.
EXAMPLE 32
Into a suitable reaction vessel equipped with a thermometer, condenser and
magnetic stirring bar were charged phosphorous acid (18-93 g) and water
(30 ml). Dissolution of the phosphorous acid was achieved by stirring and
then concentrated HCl (20 ml) was added. To the resulting solution was
added 10.0 g of N-benzylethylenediamine (Eastman Kodak Co., Rochester,
N.Y.) all at once. A heating mantel was installed and the solution heated
to reflux. Paraformaldehyde (6.64 g) was added in small portions to the
refluxing solution over a 2 hour period after which the solution was
allowed to continue refluxing for an additional 2 hours. After cooling to
room temperature the solution was concentrated under vacuum to low volume
and treated with 25 mL of concentrated HCl. After setting for 5 days the
precipitate was filtered and washed with 3N HCl. Drying of the solid gave
16.96 g (60% yield) of N-benzylethylenediaminetrimethylenephosphonic acid
(BzEDTMP) as an off-white hygroscopic solid. This product showed the
expected two singlets in a 2:1 ratio in the decoupled phosphorous-31 NMR
spectrum run on a Bruker Ac-250 MHz NMR spectrometer, pH 2.0.
.sup.31 P (D.sub.2 O): .delta. 8.20 (s, 2P), 7.37 (s, 1P)
EXAMPLE 33
A 79.4 mg portion of BzEDTMP was dissolved in 1.5 ml of HoCl.sub.3
(2.6.times.10.sup.-4 M in 0.046N HCl spiked with Ho-166 isotope) by the
addition of 50% NAOH. The final pH was 7.5 and the percent of holmium
complexed was determined by cation exchange chromatography to be greater
than 97%.
The composition of Examples 29, 31 and 33 were each injected into rats and
the biolocalization of Hol66 was determined; the data is reported in Table
X.
TABLE X
______________________________________
Example Nos.
% Dose in 29.sup.a 31.sup.a
33.sup.a
______________________________________
Skeleton 68 65 65
Blood 0.12 0.22 0.09
Liver 0.14 0.28 0.13
Muscle 0.34 0.68 0.31
Kidney 0.30 0.44 0.34
Spleen 0.01 0.01 0.01
Bone/blood 470 245 542
Bone/Muscle 1133 520 1093
______________________________________
.sup.a = represent the average of the results of 3 rats
EXAMPLE 34
A female Irish Wolfhound with a possible osteosarcoma on the left distal
radius was injected with a composition containing .sup.166 Ho-EDTMP (0.97
mCi/kg). The dog lived an additional 9 months which is greater than the
usual expected life of a dog with osteosarcoma.
COMPARATIVE EXAMPLES I
In a method similar to that previously used, compositions were prepared
containing complexes of Sm-153 with several commercially available
phosphonic acids which do not contain the alkylene linkage between the
nitrogen and the phosphorus atoms.
##STR5##
The two hour biolocalization of Sm-153 in rats for these compositions was
determined as previously described. The results are given in Table A. The
ligands used include methylendiphosphonic acid (MDP) and
hydroxyethylidinediphosphonic acid (HEDP) which contain a P-CH.sub.2
PO.sub.3 H.sub.2 and a P-C(CH.sub.3 (OH)-PO.sub.3 H.sub.2 linkage,
respectively; pyrophosphate (PYP) which contains a P-O-PO.sub.3 H.sub.2
linkage; and imidodiphosphate (IDP) which contains a N-PO.sub.3 H.sub.2
linkage. Metal complexes of these ligands are known skeletal agents. For
example, Tc complexes of MDP, HEDP, and PYP have been used commercially as
diagnostic bone agents. However, these ligands were inadequate for
selectively delivering Sm-153 to the skeletal system a3 exemplified by the
low bone to soft tissue ratios. In most cases, a large fraction of the
radioactivity was found in the liver.
Table A shows the biolocalization of Sm-153 in rats two hours after
injection and the results represent the percent of injected dose in
tissue.
TABLE A
______________________________________
Sm-153 Sm-153 Sm-153 Sm-153
% Dose In
MDP.sup.a HEDP.sup.a
PYP.sup.b
IDP.sup.b
______________________________________
Bone 2 21 2 0.6
Liver 85 3.5 73 36
Blood 0.23 13 0.23 0.04
Bone/Blood
9 1 7 11
Bone/Muscle
7 11 15 8
______________________________________
.sup.a = represent the average of the results of 5 rats
.sup.b = represent the average of the results of 3 rats
COMPARATIVE EXAMPLES II
In a method similar to that previously used, compositions were prepared
containing complexes of Ho-166 with several commercially available
phosphonic acids which do not contain the alkylene linkage between the
nitrogen and phosphorus atoms.
##STR6##
The two hour biolocalization of Ho-166 in rats for these compositions was
determined as previously described. The results are given in Table B. The
ligands used include methylenediphosphonic acid (MDP),
hydroxyethylidinediphosphonic acid (HEDP) and hydroxymethylenediphosphonic
acid (HDP) which contain a P-CH.sub.2 -PO.sub.3 H.sub.2, a P-C(CH.sub.3)
(OH)-PO.sub.3 H.sub.2 and a P-C(OH)-PO.sub.3 H.sub.2 linkage respectively;
pyrophosphate (PYP) which contains a P-O-PO.sub.3 H.sub.2 linkage; and
imidodiphosphate (IDP) which contains a N-PO.sub.3 H.sub.2 linkage. Metal
complexes of these ligands are known skeletal agents. For example, Tc
complexes of MDP, HEDP and PYP have been used commercially as diagnostic
bone agents. However, these ligands are inadequate for selectively
delivering Ho-166 to the skeletal system as exemplified by the low bone to
soft tissue ratios. In most cases, a large fraction of the radioactivity
was found in the liver.
Table B shows the biolocalization of Ho-166 in rats two hours after
injection and the results represent the percent of injected dose in
tissue.
TABLE B
______________________________________
Ho-166 Ho-166 Ho-166
Ho-166 H0-166
% Dose In MDP.sup.a
HEDP.sup.a
PYP.sup.a
IDP.sup.b
HDP.sup.b
______________________________________
Bone 1 25 3 2 0.2
Liver 64 1.8 67 65 2
Blood 1.5 10.1 0.63 0.28 0.04
Bone/Blood
0.5 2 4 6 4
Bone/Muscle
6 17 13 17 4
______________________________________
.sup.a = the average of the results of 2 rats
.sup.b = the average of the results of 1 rat
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